Hello again.
Need your help! Does anyone can suggest me kind of standart protocol for
preparing oleic acid medium, in particularly I'd like to use YPOT? I need to
use it for peroxisomes proliferation induction in wild type S.cerevisiae.
I've read a lot of articles and still can't find what I need. I've tried one
from Thieringer, where they use 0,15%(w/v)oleic acid and 0,015%(v/v)Tween
40. But I have some questions regarding:
- potassium phosphate buffer: which molarity or precentage I need to use? In
that article they suggest 30mM K2HPO4, and then to adjust pH of final medium
to 6.0, but K2HPO4 is too basic.. As I understand I need it to get right pH
only, so I tried to titrate with KH2PO4 to get pH 6. Is it correct way of
doing it?
- preparing oleic acid stock with Tween 40. Do I need to add them after
autoclaving? What's the ratio for this components ( in different papers they
use different). I did in a following way: made a stock of 150ul Tween 40 +
1.5 g oleic acid, and boil it, then added to autoclaved medium, but I'm not
sure that in this way olec acid willbe in solution. Ideally to get oleic
acid into solution you need ethanol?
Please, if you have your protocol for this or any suggestions, let me know!
Thank you in advance
--
Anna
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